Effects of Cytokine-Induced killer cells on concanavalin-a induced hepatitis in mouse

Abstract

Autoimmune hepatitis (AIH) is a chronic inflammatory disease of liver that the pathogenic mechanisms of AIH have not yet been clarified. All patients with AIH lead to cirrhosis and liver cancer. Presently the best feasible medicines of AIH need aid immunosuppressive medications and liver transplantation which have many side effects, high cost and sustained remission. Thus, immunotherapy is an alternative therapy in which cellular material is injected into a patient. Many previous studies showed cytokine induced killer (CIK) cells, T lymphocytes that have a phenotype of NK cells, have a potential to against several diseases associated with liver. The current research aims to examine protective effects of CIK cells on Concanavalin A (Con A) induced AIH in mice. In this study we use Con A because it is a generally used model for AIH in mice, the activation and recruitment of T cells to the liver. CIK cells cultured from thymus were then used to develope a protocol for generating CIK cells by adding interferon gamma (IFN-g), monoclonal antibody (mAb) against CD3 and interleukin-2 (IL-2). In this study markers show NK1.1 41.60 %, IFN-g 8.56 %, Granzyme 53.40 % and Perforin 8.31 %. Twenty-four hours before Con A injection mice were injected with CIK cells through IV (10 x 10⁶ cells per 200 µl). Mice were intravenously (IV) injected through lateral tail vein for AIH induction (25mg/kg). Lastly mice were euthanized and organs (blood and liver) were collected for biochemical test (AST, ALT) and histopathology study. In histopathology study show significant difference between control group and experimental group, hepatocellular swelling, scattered area of leukocytic infiltration, multiple necrotic area, central veins are grossly intact and no fibrotic tissue. For biochemical test we found that the average of AST level in three group are 48.33, 398.75 and 487.33 U/ml respectively while the average of AST level are 38.33, 113.75 and 199 U/ml respectively. Statistical analysis by SPSS program were used to compare between 2 groups (ConA and ConA & CIK cells group) in pathological study and biochemical test (P<0.05). In this study concluded that CIK cells does not show protective effect on Concanavalin-A indued hepatitis in mouse.